The present invention relates to propenamide derivatives having a tripeptide, Arg-Gly-Asp, as an essential unit, polymers, copolymers and salts thereof as well as a composition for inhibiting adhesion of animal cells and a composition for inhibiting coagulation and/or adhesion of blood platelets. The present invention also relates to crosslinked polymers and crosslinked copolymers of the propenamide derivatives having a tripeptide, Arg-Gly-Asp, as an essential unit, and salts thereof as well as a substrate for cultivating animal cells.
Fibronectin is a protein involved in the cell-extracellular substrate adhesion and is likewise thought to be involved in coagulation of blood platelets and the metastasis of cancer. These interactions are mediated by a series of receptors present in the cell surface region, it is confirmed that these receptors can specifically recognize an amino acid sequence: Arg-Gly-Asp of the fibronectin although the fibronectin is a macromolecule having a molecular weight of about 250,000 and there has been reported that the sequence plays an important role in the interaction between the receptors and the fibronectin (Nature, 1984,309, p. 30). Since then, there have been conducted many studies in which an oligopeptide or polypeptide having such an amino acid sequence: Arg-Gly-Asp is used.
There have been reported various studies, such as a method for inhibiting the coagulation of blood platelets by the use of various linear and cyclic oligopeptides having an Arg-Gly-Asp sequence (Polymer Preprints, Japan, 1989,38, p. 3149; Japanese Unexamined Patent Publication (hereinafter referred to as "J.P. KOKAI") No. Hei 2-174797); a method in which a peptide having an Arg-Gly-Asp sequence is used as a cell movement-inhibiting agent (J.P. KOKAI No. Hei 2-4716); and a method using, as a cell-adhesive membrane, a PMMA film on which Arg-Gly-Asp sequences are immobilized (Polymer Preprints, Japan, 1988, 37, p. 705). In addition, J.P. KOKAI Nos. Hei 1-309682 and Hei 1-305960 disclose a method which comprises peptides having Arg-Gly-Asp sequences as essential structural units covalently bonded to a polymer and the resulting product is used as a substrate for cultivating animal cells or for biological composite artificial organs and J.P. KOKAI No. Sho 64-6217 discloses a method in which a polypeptide having Arg-Gly-Asp-Ser (SEQ ID NO:5) sequences is used as a platelet protective agent for blood taken out of the body. Further, there has been known a method comprising inhibiting the metastasis of cancer by the use of an oligopeptide having Arg-Gly-Asp sequences or a polypeptide having the sequence as repeating units (Int. J. Biol. Macromol., 1989, 11, p. 23; ibid, 1989, 11, p. 226; Jpn. J. Cancer Res., 1989, 60, p. 722).
A polymer generally has various properties and functions and therefore interactions between the polymer and a living body are significantly different from those between a low-molecular substance and a living body. There have been made many studies to link a low-molecular drug or a biologically active peptide to a polymer and to control interactions between the drug or the peptide and cells or behavior of the drug or the peptide in a living body. Such applications of polymers have widely been tried to the field of life sciences such as medical polymers, polymer drugs, materials for diagnosis, bioreactors, bioaffinity materials and the like. Such polymers and use thereof are detailed in "Polymers and medical treatments", by K. Takemoto, et. al. published by Mita Publishing, "Biotechnology Series, Immobilized Enzymes", by I. Senhata published by Kodansha and "Affinity Chromatography", by M. Yamazaki, et. al. published by Kodansha.
Propenamide derivatives prepared from propenoic acid derivatives have vinyl groups and easily polymerize or copolymerize with various anionic or cationic vinyl monomers to produce various polymers or copolymers which can be used for many purposes. It has been reported that an oligopeptide having Arg-Gly-Asp as an essential unit was introduced into a water-insoluble vinyl polymer. However, there have never been known a propenamide derivative which has an oligopeptide having Arg-Gly-Asp as an essential unit in the side chain, a water-soluble polymer thereof, a water-soluble anionic or cationic copolymer thereof and a hydrogel thereof, which are expected to have increased bonding ability to a receptor and increased stability in blood.